Our research plan follows the long-range goals stated in the initial proposal. Specifically we plan to: (1) Use the protein and mRNA assays developed over the previous funding period to look at the temporal coordination of protein and mRNA synthesis throughout the yeast mitotic cell division cycle. (2) Identify authentic galactokinase and uridinyl transferase gene recombinant plasmids from the 22 candidates isolated, (using yeast transformation analysis and hybridization-arrest-translation analysis), in order to use them as physical probes to analyze complimentary mRNA species produced at various intervals of the yeast cell cycle. (3) Identify those clones from the yeast plasmid band which contain the specific genes for which mRNA assays have been developed (alcohol dehydrogenase, alpha-glucosidase, hexokinase, beta-fructosidase) with a view toward correlating the known regulatory phenotypes with a physical map of the genome obtained through sequences characterization of the cloned genes. (4) Determine as a function of time the poly(A) length of an inducible mRNA species (uridinyl transferase and galactokinase mRNAs) throughout the induction period, to test whether poly(A) length is uniform initially, and through degradation and/or repolymerization of poly(A) in the cytoplasm, becomes heterogenous. (5) Utilize the technology developed for rapid pulse labeling and isolation of mRNA species (the technique has resolved large (approximately 30S) mRNA species which are not apparent at labeling times as short as 4 minutes) to characterize the primary and processed transcription products of the genes under investigation.